ABSTRACT
Background: The endemic character of Rift Valley fever (RVF) disease points toward an interepidemic reservoir. Although not yet identified, bats and rodents may be implicated in RVF virus (RVFV) epidemiology. In this study, we investigated the putative role of Egyptian frugivorous and insectivorous bats in RVFV epidemiology in Egypt. Methods: From 2019 to 2021, 200 bats of two different species from six Egyptian governorates were tested for phleboviruses using real-time RT-PCR (rRT-PCR) and sequence analysis. Results: Screening through rRT-PCR showed evidence of the RVFV genome only in insectivorous bats. Partial sequence and phylogenetic analysis based on S and M genome segments showed that these viruses are genetically similar to those circulating (clade A) in livestock and humans during previously reported RVFV outbreaks in 1977/78 and 2003 in Egypt. Conclusions: Our molecular data suggest that the bat Pipistrellus deserti could play a role in RVFV ecology in Egypt.
Subject(s)
Chiroptera , Rift Valley Fever , Rift Valley fever virus , Animals , Egypt/epidemiology , Phylogeny , Rift Valley Fever/epidemiology , Rift Valley fever virus/geneticsABSTRACT
Bats are considered ideal reservoirs for zoonotic viruses with emerging capabilities over the past two decades and spotted evidence points out that they may play a role as a reservoir host for SARS-CoV-2. To investigate the possible role of bats as part of SARS-CoV-2 anthropozoonotic spill-over infections in Egypt, a total of 800 samples obtained from 200 Egyptian fruit bats (Rousettus aegyptiacus) were tested for SARS-CoV-2 using quantitative RT-PCR assay (RT-qPCR). RT-qPCR analysis of RNA extracted from bat tissues showed no positive results for SARS-CoV-2 nucleic acid. These findings suggest that during the study period, the Rousettus aegyptiacus bat was not a reservoir or amplifying host for SARS-CoV-2 infection in Egypt. The lack of SARS-CoV-2 nucleic acid in Egyptian fruit bats is thought to make a significant contribution to SARS-CoV-2 epidemiology.
ABSTRACT
BACKGROUND AND AIM: Highly pathogenic avian influenza H5N8 virus of clade 2.3.4.4 was newly emerged to Egypt and firstly detected in carcasses of wild birds in November 2016. This study assessed the protection efficacy and virus shedding reduction of three different inactivated avian influenza (AI) H5 (H5N1, H5N2, and H5N3) commercial vaccines against challenge with two newly emerging highly pathogenic AI virus H5N8 Egyptian isolates in specific-pathogen-free (SPF) chicks. MATERIALS AND METHODS: 10-day-old SPF chicks (n=260) were divided into 20 groups (n=13). Groups 1-5 were vaccinated through the subcutaneous route (S/C) with 0.5 mL of H5N1 vaccine, Groups 6-10 were vaccinated (S/C) with 0.5 mL of H5N2 vaccine, and Groups 11-15 were vaccinated (S/C) with 0.5 mL of H5N3 vaccine. Positive control groups (16-19) were challenged at 25 and 31 days old (2 and 3 weeks post-vaccination [PV]) using H5N8 clade 2.3.4.4 A/duck/Egypt/F13666A/2017(H5N8) and H5N8 clade 2.3.4.4 A/chicken/Egypt/18FL6/2018(H5N8). Group 20 was left non-vaccinated as a control. All vaccinated groups were divided and challenged with both viruses at 25 and 31 days of age. The viral challenge dose was 0.1 mL of 106 EID50/0.1 mL titer/chick, and it was administered oronasally. All chicks were kept in isolators for 14 days after each challenge. Sera samples were collected weekly and at 2 weeks post-challenge (PC) to detect a humoral immune response. PC mortalities were recorded daily for 10 days to calculate the protection percentages. Tracheal swabs were collected from the challenged chicks in different groups at 3, 5, 7, and 10 days PC. Kidneys and spleens were collected at 3, 5, 7, and 10 days PC and kept in formalin for histopathological examination to assess lesions and severity scores. Tracheal swabs were inoculated in 10-day-old SPF embryonated chicken eggs for virus titration and to calculate shedding levels. RESULTS: All studied vaccines displayed 70-100% protection within 10 days PC. Hemagglutination inhibition results from sera samples revealed antibody titers ranging from 0.6 to 5.4 log2 starting at 1-week PV with the highest titers at 4 weeks PV. Challenged SPF chickens exhibited a notable reduction in virus shedding, with an average of 1.5-2 log10, compared to control birds. Various histopathological lesions with different scores were detected. CONCLUSION: Our findings suggest that the inadequate virus shedding reduction and protection efficacy of studied vaccines were variable and that the type of vaccine to be used under field conditions should be reconsidered. Study of the variability between the Egyptian old emerged AI (AIV) 2017 H5N8 strains and the new emerging AIV 2018 H5N8 is required to achieve optimal protection and limit the current economic losses.
ABSTRACT
In the present study, whispovirus immediate early 1 promoter (ie-1) was used to initiate surface expression of the hemagglutinin (HA) protein of Egyptian H5N1 avian influenza virus (AIV) by using the baculovirus expression vector system. The HA gene and whispovirus ie-1 promoter sequence were synthesized as a fused expression cassette (ie1-HA) and successfully cloned into the pFastBac-1 transfer vector. The recombinant vector was transformed into DH10Bac competent cells, and the recombinant bacmid was generated via site-specific transposition. The recombinant bacmid was used for transfection of Spodoptera frugiperda (Sf-9) insect cells to construct the recombinant baculovirus and to induce expression of the HA protein of H5N1 AIV. The recombinant glycoprotein expressed in Sf-9 cells showed hemadsorption activity. Hemagglutination activity was also detected in both extra- and intracellular recombinant HAs. Both the HA and hemadsorption activities were inhibited by reference polyclonal anti-H5 sera. Significant expression of the recombinant protein was observed on the surface of infected insect cells by using immunofluorescence. SDS-PAGE analysis of the expressed protein revealed the presence of a visually distinguishable band of ~63 kDa in size, which was absent in the non-infected cell control. Western blot analysis confirmed that the distinct 63 kDa band corresponded to the recombinant HA glycoprotein of H5N1 AIV. This study reports the successful expression of the HA protein of H5N1 AIV. The expressed protein was displayed on the plasma membrane of infected insect cells under the control of whispovirus ie-1 promoter by using the baculovirus expression vector system.
